Inhibition of Leukocyte Rolling by Nitric Oxide during Sepsis Leads to Reduced Migration of Active Microbicidal Neutrophils

ABSTRACT We developed two models of sepsis with different degrees of severity, sublethal and lethal sepsis, induced by cecal ligation and puncture. Lethal sepsis induced by cecal ligation and puncture (L-CLP) resulted in failure of neutrophil migration to the infection site and high mortality. Treatment of septic animals with aminoguanidine (AG), a nitric oxide (NO) synthase inhibitor, precluded the failure of neutrophil migration and protected the animals from death. However, cytokine-induced NO synthase (iNOS)-deficient (iNOS−/−) mice subjected to L-CLP did not present neutrophil migration failure, but 100% lethality occurred. iNOS−/− mice subjected to sublethal sepsis induced by cecal ligation and puncture (SL-CLP) also suffered high mortality despite the occurrence of neutrophil migration. This apparent paradox could be explained by the lack of microbicidal activity in neutrophils of iNOS−/− mice present at the infection site due to their inability to produce NO. Notably, SL- and L-CLP iNOS−/− mice showed high bacterial numbers in exudates. The inhibition of neutrophil migration by NO is due to inhibition of a neutrophil/endothelium adhesion mechanism, since a reduction in leukocyte rolling, adhesion, and emigration was observed in L-CLP wild-type mice. These responses were prevented by AG treatment and were not observed in the iNOS−/− L-CLP group. There was no significant change in L-selectin expression in neutrophils from L-CLP mice. Thus, it seems that the decrease in leukocyte rolling is due to a defect in the expression of adhesion molecules on endothelial surfaces mediated by iNOS-derived NO. In conclusion, the results indicate that despite the importance of NO in neutrophil microbicidal activity, its generation in severe sepsis reduces neutrophil migration by inhibiting leukocyte rolling and their firm adhesion to the endothelium, in effect impairing the migration of leukocytes and consequently their fundamental role in host cell defense mechanisms.

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Inhibition of iNOS protects against the deleterious effects of sub-lethal sepsis and ethanol in the cardiorenal system

Canadian Journal of Physiology and Pharmacology ◽

10.1139/cjpp-2021-0239 ◽

2021 ◽

Author(s):

Arthur H. Sousa ◽

Gabriel T. Do Vale ◽

Jose A Nascimento ◽

Wanessa Mayumi Carvalho Awata ◽

Carla Brigagão Pacheco da Silva ◽

...

Keyword(s):

Biochemical Analysis ◽

Molecular Mechanisms ◽

Renal Cortex ◽

Cecal Ligation ◽

Selective Inhibitor ◽

Oxygen Species ◽

Cecal Ligation And Puncture ◽

Post Surgery ◽

Lethal Sepsis ◽

Pro Inflammatory Cytokines

We tested the hypothesis that ethanol would aggravate the deleterious effects of sub-lethal cecal ligation and puncture (SL-CLP) sepsis in the cardiorenal system and that inhibition of iNOS would prevent such response. Male C57BL/6 mice were treated with ethanol for 12 weeks. One hour before SL-CLP surgery, mice were treated with N6-(1-Iminoethyl)-lysine (L-NIL, 5 mg/kg, i.p), a selective inhibitor of iNOS. A second dose of L-NIL was administered 24 h after SL-CLP surgery. Mice were killed 48 h post-surgery and blood, the renal cortex and left ventricle (LV) were collected for biochemical analysis. L-NIL attenuated the increase in serum creatinine levels induced by ethanol, but not by SL-CLP. Ethanol, but not SL-CLP increased creatine kinase (CK)-MB activity and L-NIL did not prevent this response. In the renal cortex, L-NIL prevented the redox imbalance induced by ethanol and SL-CLP. Inhibition of iNOS also decreased lipoperoxidation induced by ethanol and SL-CLP in the LV. L-NIL prevented the increase of pro-inflammatory cytokines and reactive oxygen species (ROS) induced by ethanol and/or SL-CLP in the cardiorenal system, suggesting that iNOS modulated some of the molecular mechanisms that underlie the deleterious effects of both conditions in the cardiorenal system.

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A study for a role of inducible NO synthase in septic rats with cecal ligation and puncture (CLP)

The Japanese Journal of Pharmacology ◽

10.1016/s0021-5198(19)34411-7 ◽

1999 ◽

Vol 79 ◽

pp. 97

Author(s):

Masayoshi Abe ◽

Iku Okamoto ◽

Kazuhiko Shibata ◽

Keiichi Tanaka ◽

Noriyuki Sakata ◽

...

Keyword(s):

Cecal Ligation ◽

No Synthase ◽

Cecal Ligation And Puncture ◽

Inducible No Synthase

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LOW-LEVEL EXPRESSION AND LIMITED ROLE FOR THE INDUCIBLE ISOFORM OF NITRIC OXIDE SYNTHASE IN THE VASCULAR HYPOREACTIVITY AND MORTALITY ASSOCIATED WITH CECAL LIGATION AND PUNCTURE IN THE RAT

Shock ◽

10.1097/00024382-199610000-00004 ◽

1996 ◽

Vol 6 (4) ◽

pp. 248-253 ◽

Cited By ~ 30

Author(s):

Amos Vromen ◽

Marc S. Arkovitz ◽

Basilia Zingarelli ◽

Andrew L. Salzman ◽

Victor F. Garcia ◽

...

Keyword(s):

Nitric Oxide ◽

Nitric Oxide Synthase ◽

Cecal Ligation ◽

Cecal Ligation And Puncture ◽

Low Level ◽

Limited Role ◽

Vascular Hyporeactivity ◽

Oxide Synthase

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Intrahepatic STAT-3 activation and acute phase gene expression predict outcome after CLP sepsis in the rat

AJP Gastrointestinal and Liver Physiology ◽

10.1152/ajpgi.1998.275.6.g1423 ◽

1998 ◽

Vol 275 (6) ◽

pp. G1423-G1429 ◽

Cited By ~ 13

Author(s):

Kenneth M. Andrejko ◽

Jodi Chen ◽

Clifford S. Deutschman

Keyword(s):

Gene Expression ◽

Transcription Factor ◽

Acute Phase ◽

Interleukin 6 ◽

Cecal Ligation ◽

Cecal Ligation And Puncture ◽

Signal Transducer ◽

Lethal Sepsis

Interleukin-6 (IL-6) regulates hepatic acute phase responses by activating the transcription factor signal transducer and activator of transcription (STAT)-3. IL-6 also may modulate septic pathophysiology. We hypothesize that 1) STAT-3 activation and transcription of α2-macroglobulin (A2M) correlate with recovery from sepsis and 2) STAT-3 activation and A2M transcription reflect intrahepatic and not serum IL-6. Nonlethal sepsis was induced in rats by single puncture cecal ligation and puncture (CLP) and lethal sepsis via double-puncture CLP. STAT-3 activation and A2M transcription were detected at 3–72 h and intrahepatic IL-6 at 24–72 h following single-puncture CLP. All were detected only at 3–16 h following double-puncture CLP and at lower levels than following single-puncture CLP. Loss of serum and intrahepatic IL-6 activity after double-puncture CLP correlated with mortality. Neither intrahepatic nor serum IL-6 levels correlated with intrahepatic IL-6 activity. STAT-3 activation following single-puncture CLP inversely correlated with altered transcription of gluconeogenic, ketogenic, and ureagenic genes. IL-6 may have both beneficial and detrimental effects in sepsis. Fulminant sepsis may decrease the ability of hepatocytes to respond to IL-6.

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ENDOTHELIN-1 MEDIATED INDUCIBLE NITRIC OXIDE SYNTHASE GENE EXPRESSION IN THE MOUSE ILEUM AFTER CECAL LIGATION AND PUNCTURE

Shock ◽

10.1097/00024382-199506002-00039 ◽

1995 ◽

Vol 3 ◽

pp. 11-12

Author(s):

Q Qian ◽

M A Wilson ◽

M C Chou ◽

D A Spain ◽

R N Garrison

Keyword(s):

Gene Expression ◽

Nitric Oxide ◽

Nitric Oxide Synthase ◽

Inducible Nitric Oxide Synthase ◽

Cecal Ligation ◽

Cecal Ligation And Puncture ◽

Endothelin 1 ◽

Oxide Synthase ◽

Mouse Ileum ◽

Inducible Nitric Oxide

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Galectin-1-deficient mice are protected against Trypanosoma cruzi infection through altered neutrophil migration and production of reactive oxygen species and nitric oxide

10.1101/2020.09.11.294199 ◽

2020 ◽

Author(s):

Thalita Bachelli Riul ◽

Helioswilton Sales de Campos ◽

Djalma de Souza Lima-Junior ◽

Ana Elisa Caleiro Seixas Azzolini ◽

Cristina Ribeiro de Barros Cardoso ◽

...

Keyword(s):

Nitric Oxide ◽

Reactive Oxygen Species ◽

Trypanosoma Cruzi ◽

Neutrophil Migration ◽

Reactive Oxygen ◽

Oxygen Species ◽

Infection Site ◽

Neutrophil Accumulation

SUMMARYTrypanosoma cruzi is an intracellular parasite that causes Chagas disease that affects millions of people worldwide. Many cellular and molecular aspects of this neglected disease are not fully understood. Prior studies have shown that galectin-1 (Gal-1), a β-galactoside-binding protein that regulates leukocyte recruitment to the inflammatory site, and promotes T. cruzi infection, but the mechanism is unclear. Here, we report that C57BL/6 mice lacking Gal-1 (Lgals1−/−) exhibited lower parasitemia and higher survival rates than their wildtype (WT) counterparts when infected with T. cruzi Y strain. Two weeks after infection, Lgals1−/− mice displayed greater neutrophil accumulation in infection site and heart tissue than WT mice. In T. cruzi-infected Lgals1−/− mice, infiltrated neutrophils produced increased levels of reactive oxygen species (ROS), while macrophages and neutrophils produced increased levels of nitric oxide (NO), which reduced replication and viability of parasites in vitro and downregulated IL-1β production. Pharmacological inhibition of NADPH oxidase and NO synthase during early in vivo infection reversed the protective effect of Gal-1 deficiency in Lgals1−/− mice. Together, our findings demonstrate that lacking Gal-1 favors neutrophil migration to the infection site and increases production of ROS and NO, thereby controlling the early steps of T. cruzi infection by reducing parasitemia and prolonging survival of infected mice.

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Decreased exhaled nitric oxide as a marker of postinsult immune paralysis

Journal of Applied Physiology ◽

10.1152/japplphysiol.00840.2003 ◽

2004 ◽

Vol 97 (4) ◽

pp. 1188-1194 ◽

Cited By ~ 5

Author(s):

Habiba L. Attalah ◽

Stéphanie Honoré ◽

Saadia Eddahibi ◽

Elisabeth Marcos ◽

Claude-James Soussy ◽

...

Keyword(s):

Nitric Oxide ◽

Protein Expression ◽

Lung Tissue ◽

Alveolar Macrophages ◽

Neutrophil Migration ◽

No Synthase ◽

No Production ◽

Bacterial Killing ◽

Exhaled No ◽

Immune Paralysis

Nitric oxide (NO) regulates neutrophil migration and alveolar macrophage functions such as cytokine synthesis and bacterial killing, both of which are impaired in immune paralysis associated with critical illness. The aim of this study was to determine whether NO is involved in immune paralysis and whether exhaled NO measurement could help to monitor pulmonary defenses. NO production (protein expression, enzyme activity, end products, and exhaled NO measurements) was assessed in rats after cecal ligation and puncture to induce a mild peritonitis (leading to ∼20% mortality rate). An early and sustained decrease in exhaled NO was found after peritonitis (from 1 to 72 h) compared with healthy rats [median (25th–75th percentile), 1.5 parts per billion (ppb) (1.2–1.7) vs. 4.0 ppb (3.6–4.3), P < 0.05], despite increased NO synthase-2 and unchanged NO synthase-3 protein expression in lung tissue. NO synthase-2 activity was decreased in lung tissue. Nitrites and nitrates in supernatants of isolated alveolar macrophages decreased after peritonitis compared with healthy rats, and an inhibitory experiment suggested arginase overactivity in alveolar macrophages bypassing the NO substrate. Administration of the NO synthase-2 inhibitor aminoguanidine to healthy animals reproduced the decreased neutrophil migration toward alveolar spaces that was observed after peritonitis, but l-arginine administration after peritonitis failed to correct the defect of neutrophil emigration despite increasing exhaled NO compared with d-arginine administration [4.8 (3.9–5.7) vs. 1.6 (1.3–1.7) ppb, respectively, P < 0.05]. In conclusion, the decrease in exhaled NO observed after mild peritonitis could serve as a marker for lung immunodepression.

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